Roles and regulation of polyamines during HCMV infection

多胺在 HCMV 感染过程中的作用和调节

• 批准号: 10593450
• 负责人: ADAM P. GEBALLE
• 金额: $ 17.09万
• 依托单位: FRED HUTCHINSON CANCER CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-01 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10593450
• 关键词: Affect; Amino Acid Sequence; Amino Acids; Binding; Cell physiology; Cells; Cistrons; Codon Nucleotides; Cytomegalovirus; Cytomegalovirus Infections; DNA; Data; Disease; Dissection; Engineering; Ensure; Eukaryotic Cell; Eukaryotic Initiation Factors; Event; Experimental Designs; Gene Expression; Genes; Genetic; Human; Investigation; Life; Link; Medical; Messenger RNA; Metabolism; Mutation; Neurologic Deficit; Open Reading Frames; Patients; Peptides; Pharmacology; Phenotype; Physiological; Polyamines; Post-Translational Protein Processing; Production; Proline; Proteins; Putrescine; RNA; Regulation; Reproduction; Resistance; Ribosomes; Role; Site; Spermidine; Spermine; System; Testing; Translational Regulation; Translations; Viral; Viral Genes; Virus; Virus Diseases; Virus Replication; alkalinity; experimental study; fascinate; genome sequencing; human pathogen; immune system function; in utero; infant infection; inhibitor; insight; mutant; polyproline; tool; whole genome

Human cytomegalovirus (HCMV) causes life-threatening diseases in patients with poorly functioning immune systems as well as long-lasting neurological deficits in infants infected in utero. Among the many host cell factors that are needed by HCMV to replicate efficiently are small alkaline molecules called polyamines (PAs). Among their myriad activities, PAs bind to the protein synthesizing machinery in the cell. As well, they are necessary to make a protein, known as eIF5A, that is required for linking together amino acids to make certain proteins. Although it has been known for decades that the PAs are critical for HCMV to reproduce efficiently and that HCMV infection increases the intracellular concentration of PAs, the reasons why PAs are needed and the mechanisms the virus uses to ensure that there are sufficient amounts of PAs in the cells are unknown. Intriguing data emerging from studies of several host cell proteins that control the levels of PAs suggest that PAs and eIF5A might control expression of some HCMV genes, in particular a fascination one called, gpUL4. Prior studies showed that the production of gpUL4 is greatly repressed because the machinery for making it becomes trapped on the messenger RNA as a particular sequence of amino acids are being linked together. These and other results suggest that gpUL4, the function of which has been enigmatic for decades, might contribute to the control of PAs accumulation during HCMV infection. Using various HCMV mutants that affect the production of gpUL4 along with genetic and pharmacological tools for altering PA and eIF5A abundance, Aim 1 will reveal if and how PA and eIF5A alter the trapping mechanism during viral infection. Aim 2 will first test the specific hypothesis that gpUL4, alone and during HCMV infection, regulates the concentration of PAs in the cells. Another experiment will test whether HCMV can somehow adapt to reproduce well even when PA levels are low. While these studies focus on HCMV, and especially gpUL4, the results will have broad implications for understanding control of other viral and cells genes that are likely regulated by a similar mechanism.

人类巨细胞病毒（HCMV）导致功能较差的免疫系统患者以及子宫内感染的婴儿的长期神经缺陷引起威胁生命的疾病。在HCMV有效复制的许多宿主细胞因子中，称为多胺（PAS）。在其无数活性中，PA与细胞中蛋白质合成机械结合。同样，它们也必须制造一种称为EIF5A的蛋白质，这是将氨基酸链接在一起以制造某些蛋白质所必需的。尽管数十年来已经知道PAS对于HCMV有效再现至关重要，并且HCMV感染增加了PAS的细胞内浓度，但需要PAS的原因以及病毒用来确保细胞中PAS足够量的机制尚不清楚。从控制PA的水平的几种宿主细胞蛋白的研究中出现的有趣数据表明，PAS和EIF5A可能控制某些HCMV基因的表达，特别是一种称为GPUL4的诱惑。先前的研究表明，GPUL4的产生受到极大的抑制，因为将其捕获的机械被困在信使RNA上，因为特定的氨基酸序列正在连接在一起。这些和其他结果表明，GPUL4（几十年来一直是神秘的功能）可能有助于控制HCMV感染期间的PAS积累。使用影响GPUL4产生的各种HCMV突变体以及改变PA和EIF5A丰度的遗传和药理工具，AIM 1将揭示PA和EIF5A是否以及如何改变病毒感染期间的捕获机制。 AIM 2将首先检验以下特定假设：GPUL4单独和HCMV感染期间调节细胞中PA的浓度。另一个实验将测试HCMV是否可以以某种方式适应PA水平较低的情况。尽管这些研究集中于HCMV，尤其是GPUL4，但结果将对了解可能受类似机制调节的其他病毒和细胞基因的控制具有广泛的影响。